A Simple and High Quality Method for Isolation and Extraction of Total RNA of Pholiota adipose

• Published in: Asian agricultural research Vol. 10; no. 1; pp. 40 - 44
• Main Authors: Wei, Zhimin, Li, Shunguo, Liu, Meng, Xia, Xueyan, Zhao, Yu, Zhou, Hanzhang, Guo, Chengang
• Format: Journal Article
• Language: English
• Published: Cranston Wu Chu (USA-China) Science and Culture Media Corporation 01.01.2018
• Subjects: Cancer; Electrophoresis; Gene expression; Genomes; Genomics; Methods; Nitrogen; Potassium; Quality; R&D; Research & development; Ribonucleic acid; RNA
• ISSN: 1943-9903
• DOI: 10.19601/j.cnki.issn1943-9903.2018.1.010

According to the requirements of RNA-seq technology, high-quality RNA products must meet the following requirements; (i) the sample should have good integrity of RNA, and the RNA integrity number (RIN) must reach 6.8 or altove; (ii) the total content should be greater than 10 p.g (to be calculated using the Gibo green dye method) ; ( iii) the ODM/7S0 value should be in the range of 1.8-2.2; (iv) the 25S/18S RNA concentration by agarose gel electrophoresis (AGE) is about 2, no genomic pollution, no protein or impurity pollution, and no color abnormalities in the strip observation. 01 ) , indicating that the two groups of data had extremely significant differences. [...]the liquid nitrogen ground samples were veiy poor for extracting high quality total RNA of P. adipose products. 3.3 RNA quality evaluation The OD value of RNA al ihe wavelength of 260 nm and 280 nm did not fully reflect the quality. [...]using Image J software, we carried out a quantitative analysis of 25S and 18S bands in ihree lanes, ihe grayscale ralio of 25S and 18S bands in ihree lanes was 1.85, 1.88 and 1.80 respeclively , all were close lo 2, basically conforming lo ihe non-degradable eukaryolic conlenl ralio of 25S and 18S RNA. [...]the electrophoresis results proved that liquid nitrogen grinding extraction method meets the high quality standard of RNA-seq technology, and KNA quality is high. 3.3.2RNA integrity analysis. 7 and 9.3, respeclively, all higher than the quality standard 6. [...]through comprehensive analysis, the quality of RNA extracted by Trizol single-step method with liquid nitrogen grinding meets the quality requirement of RNA-seq technology, and it is the product of high quality RNA. 4Conclusions Based on ihe urgent need of researches about antitumor and other medicinal value of P. adipose from ihe Iranscriplome level, we carried out an in-depth analysis on the difficulty in RNA extraction using RNA-seq hot spot technologies.